HP Custom siRNA is provided as High-Performance–Purity (HPP) grade. HPP-grade siRNA is the optimal siRNA purification grade for efficient gene silencing at an affordable cost. HPP-grade siRNA is synthesized using proprietary synthesis and purification processes, yielding siRNA that is >90% pure. There is no need for further time-consuming and expensive HPLC or PAGE purification. The high-throughput synthesis and purification processes enable fast turnaround times and high yields. Each siRNA duplex undergoes stringent quality control including MALDI-TOF mass spectrometry analysis. HPP Grade siRNA is economically priced allowing the use of highly pure siRNA in all routine RNAi experiments.

Procedure

HP Custom siRNA provides purified, annealed, and desalted siRNA for effective gene silencing using your siRNA sequence of choice. Custom siRNA is available in individual tubes or 96-well plates. siRNA is provided as a stable duplex and there is no need to deprotect, desalt, quantify, or anneal before use. siRNA duplexes are provided as a lyophilized pellet to ensure stability during delivery. Before use, simply resuspend the lyophilized siRNA in the siRNA Suspension Buffer provided.

A wide range of modification options

HP Custom siRNA is available with a variety of fluorescent labels and other modifications. Alexa Fluor dyes provide superior performance and can be directly substituted for other common dyes. Absorbance and emission maxima for available dyes, as well as dyes that can be replaced with spectrally similar Alexa Fluor dyes, are shown in the tables below. Other fluorescent dyes available include fluorescein, rhodamine, Cyanine 570, and Cyanine 670. HP Custom siRNA may be labeled at either the 3' or the 5' end of the sense strand, and labels do not affect the biological activity of the siRNA. As well as fluorescent labels, many other backbone and base modifications are available including amino linkers, thio linkers, and biotin, dabcyl, and phosphate modifications. Special modifications are also available on request; please contact QIAGEN Technical Service.

Spectral characteristics of Alexa Fluor dyes

Dye

Absorbance maximum (nm)

Emission maximum (nm)

Preferred replacement for:

Alexa Fluor 488

495

519

Fluorescein (FITC or FAM)

Alexa Fluor 546

556

573

Tetramethylrhodamine, Cyanine 570

Alexa Fluor 555

555

565

Cyanine 570

Alexa Fluor 647

650

665

Cyanine 670

Spectral characteristics of the other available fluorescent dyes

Dye

Absorbance maximum (nm)

Emission maximum (nm)

Cyanine 570

550

570

Cyanine 670

649

670

Fluorescein (FAM)

494

518

Tetramethylrhodamine (TAMRA)

555

580

Applications

Highly pure siRNA can be used for a variety of applications, including: